Measuring and manipulating metabolic fluxes in the tumor microenvironment

测量和操纵肿瘤微环境中的代谢通量

基本信息

批准号：
10507615
负责人：
Caroline Bartman
金额：
$ 12.9万
依托单位：
PRINCETON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10507615
关键词：
Address Breast Cancer Model Breast Cancer cell line Burn injury CD8-Positive T-Lymphocytes Carbohydrates Cell Separation Cells Citric Acid Cycle Connective Tissue Cells Consumption Dichloroacetate Diet Energy Metabolism Fatty acid glycerol esters Fellowship Fibroblasts Genetic Glycolysis Goals Growth Immune Immune response In Vitro Infusion procedures Isotope Labeling Kinetics Knock-out Knowledge Lung Malignant Neoplasms Malignant neoplasm of pancreas Mammary Neoplasms Measurement Measures Metabolic Metabolism Methods Modeling Monitor Mus Myeloid Cells NADH Neoplasm Metastasis Normal tissue morphology Nutrient Oxidative Phosphorylation PDH kinase Pancreatic Adenocarcinoma Pathway interactions Pharmacology Phase Play Population Primary Neoplasm Production Proteins Role Sorting - Cell Movement Supporting Cell System Techniques Testing Therapeutic Tissues Tracer Up-Regulation cancer cell cancer therapy cell type chemotherapy drug metabolism experimental study feeding fighting genetically modified cells immune checkpoint immune checkpoint blockade in vivo ketogenic diet kinase inhibitor melanoma neoplastic cell overexpression oxidative damage pyruvate dehydrogenase stressor tumor tumor growth tumor metabolism tumor microenvironment

项目摘要

Measuring and manipulating metabolic fluxes in the tumor microenvironment Tumors have altered metabolism compared to normal tissues, which suggests that drugging metabolism could kill tumors while sparing healthy tissues. However, tumor metabolism has chiefly been measured in vitro, while recent studies have showed that tumor metabolism in the body is distinct from in vitro systems. Therefore, the field needs approaches to measure tumor metabolic fluxes in vivo. During my postdoctoral fellowship, I developed methods to measure glycolytic and tricarboxylic acid cycle (TCA) flux in vivo using kinetic infusion of isotope-labeled tracers. These approaches revealed that tumors have much lower TCA flux than healthy tissues (5 mouse tumor models examined). Though the tumors had higher glycolytic flux than healthy tissues, the total ATP production rate from glycolysis plus TCA cycle-driven oxidative phosphorylation was significantly lower in tumors than in healthy tissues. Moreover, feeding mice a high-fat ketogenic diet increased tumor TCA flux and slowed tumor growth synergistically when combined with chemotherapy. These findings raise two key questions. First, since tumors in vivo are a mix of cancer cells and other infiltrating cells, what is the metabolism of cancer cells versus immune cells or fibroblasts in tumors? Second, can directly upregulating tumor TCA flux slow tumor growth? I propose first to combine my glycolysis and TCA cycle measuring techniques with immunomagnetic and sorting strategies to measure fluxes in cancer cells, immune cells, and fibroblasts (Aim 1). I will apply this strategy to melanoma, a tumor type infiltrated by CD8 T cells which can help control the tumor, and to pancreatic adenocarcinoma, a tumor type where fibroblasts and myeloid cells can be even more abundant than cancer cells. Next, I will directly upregulate TCA flux in tumor cells by using genetic and pharmacologic approaches: overexpressing the NADH uncoupler protein mito- LbNOX, knockout of the TCA suppressor protein PDK, and inhibition of PDK with dichloroacetate. I will confirm that these strategies increase TCA flux using the method I developed and will test whether increased TCA flux slows tumor growth in primary and metastatic breast tumors. Successful completion of these aims will reveal the metabolism of different cell populations in the tumor microenvironment and will test TCA upregulation as a therapeutic strategy in cancer.

肿瘤微环境中的测量和操纵代谢通量与正常组织相比，肿瘤的新陈代谢改变了，这表明吸毒新陈代谢可能在保留健康组织的同时杀死肿瘤。但是，肿瘤代谢主要是在体外测量，而最近的研究表明，体内的肿瘤代谢不同于体外系统。因此，该领域需要在体内测量肿瘤代谢通量的方法。在我期间博士后奖学金，我开发了测量糖酵解和三羧酸周期（TCA）通量的方法使用同位素标记的示踪剂动力学输注。这些方法表明肿瘤的较低 TCA通量比健康组织（检查了5个小鼠肿瘤模型）。虽然肿瘤的糖酵解通量较高比健康组织，糖酵解和TCA周期驱动的氧化的总ATP生产率肿瘤的磷酸化明显低于健康组织。此外，给小鼠喂养高脂当与化学疗法。这些发现提出了两个关键问题。首先，由于体内肿瘤是癌细胞和其他的混合物浸润细胞，癌细胞与免疫细胞或肿瘤中成纤维细胞的代谢是什么？第二，可以直接上调肿瘤TCA通量缓慢的肿瘤生长吗？我首先建议将我的糖酵解和TCA结合通过免疫磁和分类策略测量循环测量技术，以测量癌细胞中的通量，免疫细胞和成纤维细胞（AIM 1）。我将将这种策略应用于黑色素瘤，这种肿瘤类型由CD8 T浸润可以帮助控制肿瘤的细胞，以及胰腺腺癌，一种肿瘤类型，成纤维细胞和髓样细胞比癌细胞更丰富。接下来，我将直接上调肿瘤中的TCA通量通过使用遗传和药理方法：过表达NADH解偶联剂蛋白Mito- LBNOX，TCA抑制蛋白PDK的敲除，并用二氯乙酸抑制PDK。我会确认这些策略使用我开发的方法增加了TCA通量，并将测试是否增加TCA通量减慢原发性和转移性乳腺肿瘤的肿瘤生长。这些目标的成功完成将揭示肿瘤微环境中不同细胞种群的代谢，并将TCA上调作为一个癌症的治疗策略。